GSBS Student Publications

Title

Single-Molecule Imaging Reveals that Argonaute Reshapes the Binding Properties of Its Nucleic Acid Guides

Student Author(s)

William Salomon; Samson Jolly

GSBS Program

Biochemistry & Molecular Pharmacology

UMMS Affiliation

RNA Therapeutics Institute; Department of Biochemistry and Molecular Pharmacology

Date

7-2-2015

Document Type

Article

Medical Subject Headings

Animals; Argonaute Proteins; Bacterial Proteins; Mice; MicroRNAs; Molecular Imaging; Nucleic Acid Hybridization; RNA, Guide; RNA-Induced Silencing Complex; Thermodynamics; Thermus thermophilus

Disciplines

Biochemistry | Molecular Biology

Abstract

Argonaute proteins repress gene expression and defend against foreign nucleic acids using short RNAs or DNAs to specify the correct target RNA or DNA sequence. We have developed single-molecule methods to analyze target binding and cleavage mediated by the Argonaute:guide complex, RISC. We find that both eukaryotic and prokaryotic Argonaute proteins reshape the fundamental properties of RNA:RNA, RNA:DNA, and DNA:DNA hybridization—a small RNA or DNA bound to Argonaute as a guide no longer follows the well-established rules by which oligonucleotides find, bind, and dissociate from complementary nucleic acid sequences. Argonautes distinguish substrates from targets with similar complementarity. Mouse AGO2, for example, binds tighter to miRNA targets than its RNAi cleavage product, even though the cleaved product contains more base pairs. By re-writing the rules for nucleic acid hybridization, Argonautes allow oligonucleotides to serve as specificity determinants with thermodynamic and kinetic properties more typical of RNA-binding proteins than of RNA or DNA.

Rights and Permissions

Citation: Salomon WE, Jolly SM, Moore MJ, Zamore PD, Serebrov V. Single-Molecule Imaging Reveals that Argonaute Reshapes the Binding Properties of Its Nucleic Acid Guides. Cell. 2015 Jul 2;162(1):84-95. doi: 10.1016/j.cell.2015.06.029. PubMed PMID: 26140592; PubMed Central PMCID: PMC4503223. Link to article on publisher's website

DOI of Published Version

10.1016/j.cell.2015.06.029

Related Resources

Link to article in PubMed

Journal Title

Cell

PubMed ID

26140592